Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1976 Jul;14(1):55–61. doi: 10.1128/iai.14.1.55-61.1976

Pseudomonas aeruginosa exotoxin: purification by preparative polyacrylamide gel electrophoresis and the development of a highly specific antitoxin serum.

L T Callahan 3rd
PMCID: PMC420843  PMID: 820647

Abstract

Pseudomonas aeruginosa exotoxin has been purified to a specific activity of 12,000 to 16,000 mouse median lethal doses/mg of protein. Total recovery was about 25%, and the degree of purification was approximately 3,000-fold. Preparative polyacrylamide gel electrophoresis greatly facilitated purification. As judged by analytical disc gel electrophoresis, the purified toxin contained one major band of protein and only a negligible amount of contamination. Antiserum prepared against the purified toxin neutralized the lethal activity of crude toxin preprations and reacted by double immunodiffusion with a single component of concentrated broth cultures of P. aeruginosa isolates obtained from a clinical source.

Full text

PDF
55

Images in this article

59Image
on p.59
59Image
on p.59

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atik M., Liu P. V., Hanson B. A., Amini S., Rosenberg C. F. Pseudomonas exotoxin shock. A preliminary report of studies in dogs. JAMA. 1968 Jul 15;205(3):134–140. doi: 10.1001/jama.205.3.134. [DOI] [PubMed] [Google Scholar]
  2. Callahan L. T., 3rd Purification and characterization of Pseudomonas aeruginosa exotoxin. Infect Immun. 1974 Jan;9(1):113–118. doi: 10.1128/iai.9.1.113-118.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chrambach A., Reisfeld R. A., Wyckoff M., Zaccari J. A procedure for rapid and sensitive staining of protein fractionated by polyacrylamide gel electrophoresis. Anal Biochem. 1967 Jul;20(1):150–154. doi: 10.1016/0003-2697(67)90272-2. [DOI] [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. Fink M. A., Feller W. F., Sibal L. R. Methods for detection of antibody to the mammary tumor virus. J Natl Cancer Inst. 1968 Dec;41(6):1395–1400. [PubMed] [Google Scholar]
  6. Genell S., Heideman M., Leandoer L., Ljungqvist U. Platelet and fibrinogen reactions in Pseudomonas aeruginosa exotoxin shock. Eur Surg Res. 1973;5(5):321–332. doi: 10.1159/000127673. [DOI] [PubMed] [Google Scholar]
  7. Iglewski B. H., Kabat D. NAD-dependent inhibition of protein synthesis by Pseudomonas aeruginosa toxin,. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2284–2288. doi: 10.1073/pnas.72.6.2284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Knudsen R. C., Callahan L. T., 3rd, Ahmed A., Sell K. W. Use of microculture plates and the multiple automated sample harvester for in vitro microassay of bacterial toxins. Appl Microbiol. 1974 Aug;28(2):326–327. doi: 10.1128/am.28.2.326-327.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Liu P. V. Exotoxins of Pseudomonas aeruginosa. I. Factors that influence the production of exotoxin A. J Infect Dis. 1973 Oct;128(4):506–513. doi: 10.1093/infdis/128.4.506. [DOI] [PubMed] [Google Scholar]
  11. Liu P. V., Hsieh H. Exotoxins of Pseudomonas aeruginosa. 3. Characteristics of antitoxin A. J Infect Dis. 1973 Oct;128(4):520–526. doi: 10.1093/infdis/128.4.520. [DOI] [PubMed] [Google Scholar]
  12. Liu P. V. The roles of various fractions of Pseudomonas aeruginosa in its pathogenesis. 3. Identity of the lethal toxins produced in vitro and in vivo. J Infect Dis. 1966 Oct;116(4):481–489. doi: 10.1093/infdis/116.4.481. [DOI] [PubMed] [Google Scholar]
  13. Liu P. V. The roles of various fractions of Pseudomonas aeruginosa in its pathogenesis. II. Effects of lecithinase and protease. J Infect Dis. 1966 Feb;116(1):112–116. doi: 10.1093/infdis/116.1.112. [DOI] [PubMed] [Google Scholar]
  14. Liu P. V., Yoshii S., Hsieh H. Exotoxins of Pseudomonas aeruginosa. II. Concentration, purification, and characterization of exotoxin A. J Infect Dis. 1973 Oct;128(4):514–519. doi: 10.1093/infdis/128.4.514. [DOI] [PubMed] [Google Scholar]
  15. MCDONALD C. E., CHEN L. L. THE LOWRY MODIFICATION OF THE FOLIN REAGENT FOR DETERMINATION OF PROTEINASE ACTIVITY. Anal Biochem. 1965 Jan;10:175–177. doi: 10.1016/0003-2697(65)90255-1. [DOI] [PubMed] [Google Scholar]
  16. Pavlovskis O. R., Gordon F. B. Pseudomonas aeruginosa exotoxin: effect on cell cultures. J Infect Dis. 1972 Jun;125(6):631–636. doi: 10.1093/infdis/125.6.631. [DOI] [PubMed] [Google Scholar]
  17. Pavlovskis O. R. Pseudomonas aeruginosa exotoxin: effect on cellular and mitochondrial respiration. J Infect Dis. 1972 Jul;126(1):48–53. doi: 10.1093/infdis/126.1.48. [DOI] [PubMed] [Google Scholar]
  18. Pavlovskis O. R., Shackelford A. H. Pseudomonas aeruginosa exotoxin in mice: localization and effect on protein synthesis. Infect Immun. 1974 Mar;9(3):540–546. doi: 10.1128/iai.9.3.540-546.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES